Cargando…

Multi-phosphorylation reaction and clustering tune Pom1 gradient mid-cell levels according to cell size

Protein concentration gradients pattern developing organisms and single cells. In Schizosaccharomyces pombe rod-shaped cells, Pom1 kinase forms gradients with maxima at cell poles. Pom1 controls the timing of mitotic entry by inhibiting Cdr2, which forms stable membrane-associated nodes at mid-cell....

Descripción completa

Detalles Bibliográficos
Autores principales: Gerganova, Veneta, Floderer, Charlotte, Archetti, Anna, Michon, Laetitia, Carlini, Lina, Reichler, Thais, Manley, Suliana, Martin, Sophie G
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6555594/
https://www.ncbi.nlm.nih.gov/pubmed/31050340
http://dx.doi.org/10.7554/eLife.45983
_version_ 1783425183154962432
author Gerganova, Veneta
Floderer, Charlotte
Archetti, Anna
Michon, Laetitia
Carlini, Lina
Reichler, Thais
Manley, Suliana
Martin, Sophie G
author_facet Gerganova, Veneta
Floderer, Charlotte
Archetti, Anna
Michon, Laetitia
Carlini, Lina
Reichler, Thais
Manley, Suliana
Martin, Sophie G
author_sort Gerganova, Veneta
collection PubMed
description Protein concentration gradients pattern developing organisms and single cells. In Schizosaccharomyces pombe rod-shaped cells, Pom1 kinase forms gradients with maxima at cell poles. Pom1 controls the timing of mitotic entry by inhibiting Cdr2, which forms stable membrane-associated nodes at mid-cell. Pom1 gradients rely on membrane association regulated by a phosphorylation-dephosphorylation cycle and lateral diffusion modulated by clustering. Using quantitative PALM imaging, we find individual Pom1 molecules bind the membrane too transiently to diffuse from pole to mid-cell. Instead, we propose they exchange within longer lived clusters forming the functional gradient unit. An allelic series blocking auto-phosphorylation shows that multi-phosphorylation shapes and buffers the gradient to control mid-cell levels, which represent the critical Cdr2-regulating pool. TIRF imaging of this cortical pool demonstrates more Pom1 overlaps with Cdr2 in short than long cells, consistent with Pom1 inhibition of Cdr2 decreasing with cell growth. Thus, the gradients modulate Pom1 mid-cell levels according to cell size.
format Online
Article
Text
id pubmed-6555594
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher eLife Sciences Publications, Ltd
record_format MEDLINE/PubMed
spelling pubmed-65555942019-06-12 Multi-phosphorylation reaction and clustering tune Pom1 gradient mid-cell levels according to cell size Gerganova, Veneta Floderer, Charlotte Archetti, Anna Michon, Laetitia Carlini, Lina Reichler, Thais Manley, Suliana Martin, Sophie G eLife Cell Biology Protein concentration gradients pattern developing organisms and single cells. In Schizosaccharomyces pombe rod-shaped cells, Pom1 kinase forms gradients with maxima at cell poles. Pom1 controls the timing of mitotic entry by inhibiting Cdr2, which forms stable membrane-associated nodes at mid-cell. Pom1 gradients rely on membrane association regulated by a phosphorylation-dephosphorylation cycle and lateral diffusion modulated by clustering. Using quantitative PALM imaging, we find individual Pom1 molecules bind the membrane too transiently to diffuse from pole to mid-cell. Instead, we propose they exchange within longer lived clusters forming the functional gradient unit. An allelic series blocking auto-phosphorylation shows that multi-phosphorylation shapes and buffers the gradient to control mid-cell levels, which represent the critical Cdr2-regulating pool. TIRF imaging of this cortical pool demonstrates more Pom1 overlaps with Cdr2 in short than long cells, consistent with Pom1 inhibition of Cdr2 decreasing with cell growth. Thus, the gradients modulate Pom1 mid-cell levels according to cell size. eLife Sciences Publications, Ltd 2019-05-03 /pmc/articles/PMC6555594/ /pubmed/31050340 http://dx.doi.org/10.7554/eLife.45983 Text en © 2019, Gerganova et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Cell Biology
Gerganova, Veneta
Floderer, Charlotte
Archetti, Anna
Michon, Laetitia
Carlini, Lina
Reichler, Thais
Manley, Suliana
Martin, Sophie G
Multi-phosphorylation reaction and clustering tune Pom1 gradient mid-cell levels according to cell size
title Multi-phosphorylation reaction and clustering tune Pom1 gradient mid-cell levels according to cell size
title_full Multi-phosphorylation reaction and clustering tune Pom1 gradient mid-cell levels according to cell size
title_fullStr Multi-phosphorylation reaction and clustering tune Pom1 gradient mid-cell levels according to cell size
title_full_unstemmed Multi-phosphorylation reaction and clustering tune Pom1 gradient mid-cell levels according to cell size
title_short Multi-phosphorylation reaction and clustering tune Pom1 gradient mid-cell levels according to cell size
title_sort multi-phosphorylation reaction and clustering tune pom1 gradient mid-cell levels according to cell size
topic Cell Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6555594/
https://www.ncbi.nlm.nih.gov/pubmed/31050340
http://dx.doi.org/10.7554/eLife.45983
work_keys_str_mv AT gerganovaveneta multiphosphorylationreactionandclusteringtunepom1gradientmidcelllevelsaccordingtocellsize
AT floderercharlotte multiphosphorylationreactionandclusteringtunepom1gradientmidcelllevelsaccordingtocellsize
AT archettianna multiphosphorylationreactionandclusteringtunepom1gradientmidcelllevelsaccordingtocellsize
AT michonlaetitia multiphosphorylationreactionandclusteringtunepom1gradientmidcelllevelsaccordingtocellsize
AT carlinilina multiphosphorylationreactionandclusteringtunepom1gradientmidcelllevelsaccordingtocellsize
AT reichlerthais multiphosphorylationreactionandclusteringtunepom1gradientmidcelllevelsaccordingtocellsize
AT manleysuliana multiphosphorylationreactionandclusteringtunepom1gradientmidcelllevelsaccordingtocellsize
AT martinsophieg multiphosphorylationreactionandclusteringtunepom1gradientmidcelllevelsaccordingtocellsize